Data underlying the publication: Theoretical Minimum Uncertainty of Single-Molecule Localizations Using a Single-Photon Avalanche Diode Array

The objective of this research was to find the theoretical minimum uncertainty for single-molecule localizations using a single-photon avalanche diode (SPAD). The key questions here was whether or not the binary output of a SPAD impacts the theoretical minimum uncertainty for the SPAD image model, or whether it is identical to the previously derived theoretical minimum uncertainty for an image model using Poissonian photon statistics. <br>After having theoretically derived this minimum uncertainty and having verified this result using simulation data we concluded that the difference between the Poissonian and binary image model is largest when saturation occurs in the pixels. To test this we had to create saturation in an experimental setup. <br>To do this we have built a TIRF setup where we observed DNA-PAINT nanorulers using a SwissSPAD2. Using this setup we acquired one large dataset taken at the minimum possible exposure time of the SwissSPAD2: 10 us. We then used this dataset to aggregate multiple frames into a single image, thereby setting any pixel value above 1 to 1, hereby creating saturation. <br>The .tiff data in this data set shows images for 8 different numbers of aggregated frames, from 16 aggregated frames to 2048 aggregated frames.<br>The two zipped directories contain emitter location data that is obtained when applying a localization pipeline on the .tiff files. Using the files in these directories allow for immediate reproduction of the plots in the paper.

本研究的目标是探究使用单光子雪崩二极管（single-photon avalanche diode, SPAD）实现单分子定位时的理论最小不确定度。本研究的核心问题在于：SPAD的二进制输出是否会影响SPAD成像模型的理论最小不确定度，亦或是其与此前基于泊松光子统计推导得到的成像模型理论最小不确定度完全一致？
在完成该最小不确定度的理论推导，并通过仿真数据验证结果后，我们得出结论：当像素发生饱和时，泊松成像模型与二进制成像模型之间的差异最为显著。为验证这一结论，我们需要在实验装置中实现像素饱和。
为此，我们搭建了全内反射荧光显微镜（total internal reflection fluorescence microscope, TIRF）实验装置，使用SwissSPAD2观测DNA-PAINT纳米标尺。利用该装置，我们以SwissSPAD2的最小可行曝光时间10微秒采集了一组大型数据集。随后，我们将该数据集中的多帧图像聚合为单幅图像，并将所有像素值大于1的像素统一置为1，以此实现像素饱和。
本数据集内的.tiff格式图像涵盖8种不同的聚合帧数场景，聚合帧数范围为16帧至2048帧。
两个压缩目录中包含了对上述.tiff文件应用定位算法流程后得到的发射器定位数据。使用该目录内的文件，可直接复现论文中的相关绘图。